A comparative study of VITEK-2, Double Disc Synergy and Combined Disc Methods for detection of ESBL (Extended Spectrum Beta-Lactamase) production in Escherichia coli and Klebsiella pneumoniae strains

Abstract

Aim: In this prospective study we aimed to compare the effectiveness of VITEK-2 (bioMérieux, France) automated system, double-disc synergy (DDST) versus combined disc test (CDT) in detecting the Extended-Spectrum Beta-Lactamase (ESBL) positivity in Escherichia coli and Klebsiella pneumoniae strains isolated from various clinical samples. Material and Method: E.coli and K. pneumoniae strains inoculated on Mueller Hinton Agar plate. Susceptibility tests were performed with the VITEK 2 (BioMérieux, France) system before. Afterward, EBSL positivity was investigated manually DDST and CDT. Minimal inhibitor concentration (MIC) results of three tests were compared with each other according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. Results: 184 E. coli and K.pneumoniae strains, elevated. 92.9% of 98 patients with VITEK 2 positive results were positive with combined disc and DDS method, 100% of the 86 patients with negative results of VITEK 2 were negative with combined disc and MDC. Conclusion: VITEK 2 was found to have a sensitivity of 100%, a specificity of 92.4%, a positive predictive value of 92.8% and a negative predictive value of 100%. VITEK 2 was found to be compatible with validation tests for ESBL positivity.

Keywords

• Escherichia coli
• Klebsiella pneumoniae
• VITEK-2
• Double Disk Synergy (DDS) Test
• Extended-Spectrum Beta-Lactamase
• Combined disc test

References

1. 1. Forbes A, Sahm F, Weissfeld S. Principles of antimicrobial action and resistance, In: Fabiano K, Lester S, Wurm E(eds), Bailey & Scott’s Diagnostic Microbiology. 2003,11thed. St. Louis. CV Mosby Co, Missouri. p: 214-220.
2. 2. Livermore DM.Defining an extended-spectrum beta-lactamase. Clin Microbiol Infect. 2008;14 (1): 3-10.
3. 3. Livermore DM. Beta-lactamases in laboratory and clinical resistance. Clin Microbiol Rev. 1995; 8(4): 557-584.
4. 4. Albertini M, Benoit C, Berardi L. Surveillance of methicillin-resistant Staphylococcus aureus (MRSA) and Enterobacteriaceae producing extended spectrum beta-lactamase (ESBL) in Northern France: a five-year multicentre incidence study. J Hosp Infect. 2002; 52(2):107-113.
5. 6. Kang C, Kim SH, Park WB, et al. Bloodstream infections due to extended spectrum beta-lactamases producing E.coli and K.pneumoniae: Risk factors for treatment outcome with special emphasis on antimicrobial. Antimicrobial Agents Chemother. 2004; 48(12): 4574-4581.
6. 7. Ramphal R, Ambrose PG. Extended spectrum beta-lactamases and clinical outcomes: Current data. Clin Inf Dis. 2006; 42 Suppl 4: 164-172.
7. 8. Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: special needs for fastidious organisms and difficult-to-detect resistance mechanisms. Clin Infect Dis. 2000; 30(5): 799-808.
8. 9. Clinical and Laboratory Standards Institute. Performance Standarts for Antimicrobial Susceptibility Testing. 24th Informational Supplement. CLSI Document M100-S24, CLSI, Wayne, PA, 2014.

9. 10. Mehli M, Zer Y, Gayyurhan E. Çeşitli klinik örneklerden izole edilen Enterobacteriaceae suşlarında GSBL oluşturmanın ÇDST ve VITEK 2 yöntemleri ile araştırılması. ANKEM Derg. 2007; 21(3):71-75.
10. 11. Fincancı M, Ulutürk R, Eren G, et al. Klebsiella pneumoniae, Klebsiella oxytoca ve Escherichia coli kökenlerinde genişletilmiş spektrumlu beta laktamazların araştırılmasında kullanılan çeşitli yöntemlerin karşılaştırılması. İnfeksiyon Derg. 2003;17(1):55-60.
11. 12. Öztürk CE, Kaya DA, Yücel M, , et al.. Genişlemiş Spektrumlu Beta-Laktamaz Tespitinde Kullanılan Bazı Fenotipik Testlerin Karşılaştırılması. ANKEM Derg. 2010;24(3):111-116.
12. 13. Yavuz MT, Ersan G, Süvarierel M. Enterobacteriaceae kökenlerinde genişlemiş spektrumlu beta-laktamaz üretiminin iki farklı yöntemle araştırılması, Düzce Tıp Fak Derg. 2005;2:10-13.
13. 14. Yurtman AN, Hoşgör-Limoncu M, Ermertcan Ş, et al. Genişlemiş spektrumlu beta-laktamazların saptanmasında fenotipik yöntemlerin karşılaştırılması. İnfeksiyon Derg. 2009;23(1):5-8.
14. 15. Akçam FZ, Gönen İ, Kaya O, et al. Hastane infeksiyonu etkeni çeşitli Gram-negatif bakterilerde genişlemiş spektrumlu beta-laktamaz yapımının iki yöntemle araştırılması. Klimik Derg. 2004;17(1):47-49.
15. 16. Genç S, Dündar D. Escherichia coli ve Klebsiella pneumoniae Suşlarında GSBL Üretiminin Saptanmasında VITEK- 2 Otomatize Sistemi ile Çift Disk Sinerji Testinin Karşılaştırılması. Türk Mikrobiyol Cem Derg. 2015; 45(1):36-40.
16. 17. Ece G. Comparison of conventional method and automatized vitek system in the detection of extended spectrum beta-lactamase in Escherichia coli and Klebsiella pneumoniae isolates. Afr J Microbiol Res. 2013; 7(32):4091-95.
17. 18. Robin F, Delmas J, Schweitzer C, et al. Evaluation of the Vitek-2 extended-spectrum β-lactamase test against non-duplicate strains of Enterobacteriaceae producing a broad diversity of well-characte-rised β-lactamases . Clin Microbiol Infect. 2008; 14(2):148-154.
18. 19. Spanu T, Sanguinetti M, Tumbarello M, et al. Evaluation of the new VITEK 2 extended-spectrum beta-lactamase (ESBL) test for rapid detection of ESBL production in Enterobacteriaceae isolates. J Clin Microbiol. 2006; 44(9):3257-3262.
19. 20. Kaçmaz B, Ece G. Genişlemiş spektrumlu betalaktamaz saptanmasında ikinci, üçüncü ve dördüncü kuşak sefalosporinlerin çift disk sinerji testinde kullanılması ve sefoksitin duyarlılığı. ANKEM Derg. 2010; 24(2):61-64.
20. 21. Singh RM, Singh HL. Comparative evaluation of six phenotypic methods for detecting extended-spectrum beta-lactamase-producing Enterobacteriaceae. J Infect Dev .Ctries 2014;8(4):408-415.
21. 22. Alışkan HE, Çolakoğlu Ş, Turunç T, Demiroğlu YZ. GSBL Pozitif Enterobacteriaceae ve Vankomisine Dirençli Enterokokların İdrar Kültürlerinden Erken Tespitinde ChromID ESBL Agarın Değerlendirilmesi. Mikrobiyol Bul. 2012; 46(1): 17-25.